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PubMed1996

A prospective study on the predictive value of normal sperm morphology as evaluated by computer (IVOS).

OBJECTIVE: To evaluate the IVOS (Hamilton Thorne Research Version 2.1 Dimension Program, Beverly, MA) system's ability to predict fertilization in vitro in a prospective study. DESIGN: Hospital-based academic ART program. PATIENTS: Eighty patients from the IVF-GIFT program were evaluated. The same semen sample was analyzed on a day-to-day basis by both laboratory (manual method) and the computerized system for percentage normal morphology, concentration/mL, motility, and forward progression. Only patients with two or more metaphase II (MII) oocytes available were allowed into the study and excluded where the male partner had antisperm antibodies or qualified for intracytoplasmic sperm injection (<500,000 motile spermatozoa obtained after glass wool separation). STATISTICAL ANALYSIS: Logistic regression analysis was used to study predictors of fertilization in vitro. RESULTS: Three hundred thirty-eight oocytes were obtained from 80 patients of which 239 fertilized. The logistic regression analysis of the manual method (percentage normal morphology) and IVOS indicated that both were predictors of fertilization. Sperm morphology as evaluated by IVOS in patients with <10 x 10(6) motile spermatozoa/mL retrieved after swim-up was a significant predictor of fertilization as was the number of oocytes obtained. Thus, the more oocytes obtained in the lower morphological groups, the better the chance of fertilization. The fertilization rate in the morphology group of 0% to 4% normal forms was 45.6% (37/81) and in the group >14% normal forms was 85.2% (69/81). CONCLUSIONS: It was shown that in patients where </= 10 x 10(6) motile spermatozoa were obtained, the role of morphology (evaluated by IVOS) as well as the number of oocytes were important predictors of fertilization. The computer can assist to identify these patients with a poor prognosis for fertilization.

Computer Simulation↗
PubMed1991

Sperm morphology and IVF pregnancy rate: comparison between Percoll gradient centrifugation and swim-up procedures.

Many groups currently use two methods for the separation of motile spermatozoa, swim-up (S-up) and centrifugation on discontinuous Percoll gradient (PGC), and comparison of results indicates that PGC is superior. In this study we have attempted to identify the factors explaining this difference. This laboratory has long-standing expertise in seminology, thus the parameters of sperm morphology were the obvious first choice for detailed study. First, the respective effects of S-up and PGC on sperm morphology were analysed in different types of ejaculates: 62 semen samples with normal parameters and 41 with poor parameters. Both separation techniques resulted in improved morphology in the final preparation but only the increase of morphologically normal spermatozoa in the final Percoll suspension was significant. Second, application of these techniques in our in-vitro fertilization (IVF) programme revealed that, together with the improvement of sperm morphology, a higher pregnancy rate was obtained after PGC. The ongoing pregnancy rates per oocyte retrieval were 21.1% for the S-up technique and 33.3% for the PGC technique. These data show that spermatozoa selected by PGC present an improved morphology which we believe to be linked to improvement of the quality of the in-vitro fertilized embryos and ultimately the percentage of successful IVF results.

Adult↗
PubMed1995

Recurrent failure in polar body formation and premature chromosome condensation in oocytes from a human patient: indicators of asynchrony in nuclear and cytoplasmic maturation.

All oocytes from a patient who had undergone four unsuccessful in-vitro fertilization attempts showed neither a polar body nor pronuclei when examined for fertilization. In 19 inseminated oocytes that were spread for karyotypic analysis, one haploid set of metaphase II chromosomes and a remarkable condensed structure were found. Hormonal and morphological criteria implied that the oocytes had been mature at the time of retrieval. Since non-inseminated oocytes contained only one set of metaphase II chromosomes, the condensed structure appeared to represent the sperm chromatin in the state of premature chromosome condensation due to a block in oocyte maturation. Since the first and second polar body, as well as their chromatin, were undetectable in all the patient's oocytes, a rapid maturation to metaphase II before retrieval and prolonged arrest in this state before fertilization, accompanied by degeneration of the first polar body, appear to be responsible for the condition. In accordance with this notion, degenerate spindles (typical of post-ovulatory aged oocytes) and separating chromosomes (probably representing presegregating chromatids) were observed by antitubulin immunofluorescence.

Adult↗
PubMed1995

Factors of importance for the establishment of a successful program of intracytoplasmic sperm injection treatment for male infertility.

OBJECTIVE: To establish an intracytoplasmic sperm injection treatment program for couples with male infertility and to determine those factors important for success. DESIGN: A retrospective analysis of 171 consecutive cycles of intracytoplasmic sperm injection concerning 145 infertile couples. SETTING: Infertility clinic in a private hospital associated with a university hospital. PATIENTS: Couples with infertility in the male partner whose sperm parameters were unacceptable for conventional IVF or in whom fertilization by conventional IVF failed repeatedly. INTERVENTIONS: One hundred seventy-one transvaginal oocyte retrievals were completed after superovulation with GnRH agonist and gonadotropins. MAIN OUTCOME MEASURES: The parameters evaluated included fertilization, cleavage, implantation, pregnancy, and spontaneous abortion in relation to patient indications and improved procedures. RESULTS: After intracytoplasmic sperm injection, normal fertilization occurred in 45% of the oocytes (n = 1,499). Of 171 treatment cycles, 93% of the couples had fertilization and 86% had ET. Thirty-six pregnancies were achieved. During the period studied, the mean fertilization rate increased from 21.3% during the first 17 weeks to 67.8% during the last 13 weeks, and the pregnancy rate (PR) per started cycle increased from 12.8% to 31.3%. CONCLUSIONS: Technical factors critical for achieving high rates of fertilization and pregnancy were the use of standardized intracytoplasmic sperm injection pipettes, the immobilization of sperm before injection, and the aspiration of a minimal amount of ooplasm before reinjection with the sperm. Intracytoplasmic sperm injection appears to be superior to other micromanipulation methods for alleviating male infertility.

Adult↗
PubMed1991

Male infertility: the focus shifts to sperm manipulation.

Few systemic approaches for treatment of the infertile male are available. Therefore, the focus has shifted to studies of sperm maturation in the female tract, tests of sperm function, and assisted reproduction. New methods of sperm evaluation allow a better determination of those samples that will and will not fertilize ova. These methods include the strict morphology examination, biochemical approaches such as evaluation of creatine kinase levels, and the hemizona assay. Computer assisted semen analysis has not yet proved important for diagnosis but has provided important research information. For example, the drug pentoxifylline significantly increases velocity and has also been shown to enhance in vitro fertilization. The term subfertile is extensively used in the literature but its clear definition and the prognosis for subfertile men have not been established. Approaches that may increase fertilization rates include reducing the gamete culture volume and removal of cumulus oophorus. Assisted reproductive technologies for patients with ejaculatory dysfunction have also been expanded. The use of improved semen processing techniques, advanced oocyte retrieval, and well-timed intrauterine insemination are enabling physicians to use decreasing numbers of viable sperm to achieve pregnancies.

Humans↗
PubMed1997

Pregnancy after intracytoplasmic injection of immotile sperm. A case report.

BACKGROUND: With the advent of intracytoplasmic sperm injection (ICSI), fertilization and pregnancy have been reported in complete asthenozoospermia. The major problem with the use of immotile sperm for ICSI has been differentiating between live and dead sperm. We successfully selected live sperms with the hypoosmotic swelling test, and a twin pregnancy resulted after intracytoplasmic injection of completely immotile sperm. CASE: A 32-year-old woman and her 36-year-old husband had had primary infertility for four years. Evaluation of the couple showed complete asthenozoospermia of the husband. Ovulation in the wife was successfully stimulated with clomiphene citrate, follicle-stimulating hormone and human menopausal gonadotropin. We selected the viable sperm with the hypoosmotic swelling test. ICSI was then performed. After assisted hatching, we transferred four embryos on the third day after oocyte retrieval. Intrauterine twin pregnancy was confirmed by ultrasonography four weeks later. The healthy infants, free of anomalies, were born on the 36th gestational week. CONCLUSION: The hypoosmotic swelling test is a simple method of selecting immotile and viable sperm for use in ICSI.

Adult↗
PubMed2005

Human sperm can fertilize ova inside the Graafian follicles before ovulation.

OBJECTIVE: To report an incidental finding of fertilization of ova inside the ovarian follicles before ovulation. DESIGN: Case report. SETTING: IVF Center, King Fahd Specialist Hospital, Buraidah, Al-Qassim, Saudi Arabia. PATIENT(S): One 31-year-old woman with 8 years of primary infertility. INTERVENTION(S): Long-protocol superovulation and ovum pickup (OPU). MAIN OUTCOME MEASURE(S): The retrieved nine oocytes were incubated and followed up closely. RESULT(S): Three hours after OPU and only after decoronization of oocytes, sperm were seen attached to the zonae pellucidae of six mature oocytes. Three of them were showing signs of fertilization (two pronuclei). Neither conventional IVF nor intracytoplasmic sperm injection was done, and we decided to continue incubation and observation of all oocytes. From the nine total oocytes, fertilization was demonstrated in six with attached sperm, of which four divided. On day 2 after OPU, three embryos were transferred into the uterus. CONCLUSION(S): Human sperm can migrate and ascend through the female genital tract to get through the ovarian tissue. Moreover, under certain conditions, it can penetrate the wall of an intact ovarian follicle to reach the ovum and fertilize it just before ovulation.

Adult↗
PubMed2000

Correlation of oocyte morphology with fertilization rate and embryo quality after intracytoplasmic sperm injection.

To determine whether the morphology of the oocyte assessed under light microscopy is related to the results of intracytoplasmic sperm injection (ICSI). 135 ICSI oocytes from 15 patients were analyzed. Transvaginal ultrasound-guided oocytes retrieval with oocyte morphology evaluation based on shape of the first polar body and perivitelline space followed by ICSI. After 48 hours, embryo quality was evaluated and compared to each pre-injected oocyte morphology. Normal fertilization was achieved in 81.5 per cent of the oocytes (110/135). Abnormal fertilization (3 pronuclei) was 1.5 per cent (2/135). Fertilization rate of oocytes with good morphology was higher than those with poor morphology, but there was no statistical significance (82.4% vs 79.5%; p > 0.05). Oocytes with good morphology were significantly fertilized to be embryos with good quality (p < 0.001). This study suggests that oocyte morphology correlates with embryo quality after ICSI.

Adult↗
PubMed1998

Intracytoplasmic sperm injection today: a personal review.

There no longer seem to be any categories of male factor infertility that cannot be treated with intracytoplasmic sperm injection (ICSI). Even for men with azoospermia caused either by obstruction or by germinal failure, ICSI may be performed successfully. The only failures will be in azoospermic men who have neither spermatozoa nor spermatids retrievable from the testis, but these men comprise a small percentage of the cases with severe male factor. The source of the spermatozoa and the cause of the sperm defect appear to have no effect on the success of the procedure, whether the spermatozoon is epididymal, fresh or frozen, testicular, ejaculated, or from the testicles of men with severe defects in spermatogenesis. Maturation arrest, Sertoli cell-only, cryptorchidism, chemotherapy and mumps do not appear to have a major impact on the pregnancy rate. Of all the factors studied in couples where the male is severely infertile or azoospermic, the only factor that seems to matter (as long as spermatozoa are retrieved) is the age of the wife and, to a considerably lesser extent, her ovarian reserve. Extensive genetic and paediatric follow-up studies of ICSI pregnancies have revealed no increased risk of congenital malformation (2.6%), no increased risk of de-novo autosomal abnormalities, and a 1.0% risk of sex chromosomal abnormalities. These results are very reassuring, but point to the need for careful counselling of couples with male infertility.

Cystic Fibrosis↗
PubMedDate not supplied

Maturation of monkey spermatozoa in the epididymis with respect to their ability to undergo the acrosome reaction.

Spermatozoa obtained along the length of the epididymides from five macaque monkeys (Macaca fascicularis) were examined for their ability to undergo the acrosome reaction under basal conditions and when stimulated with the calcium ionophore A23187. Under basal conditions (2.5 hours incubation in potentially capacitating medium) few epididymal sperm cells displayed the loss of acrosome as judged by staining with peanut agglutinin lectin that binds to the outer acrosomal membrane, regardless of the epididymal region from which they were retrieved. By contrast, there was a marked difference in response to short (0.5 hour) incubation with the calcium ionophore A23187, which induced acrosome reactions in the majority of caudal sperm: caput sperm failed to respond at all and corpus sperm were about 50% as responsive as caudal spermatozoa. There is thus a development of the ability to respond to the ionophore upon maturation. Ejaculated spermatozoa from the same monkeys displayed a higher rate of basal acrosomal loss but a slightly lower stimulated response than mature epididymal spermatozoa. With these maturational aspects, monkey epididymal sperm can serve as a model for the study of human sperm maturation.

Acrosome↗
PubMed1994

The use of recombinant FSH for ovulation induction.

Recombinant human FSH (recFSH) was tested in a phase II study in 50 couples superovulated for IVF. Rec FSH alone was used in 10 women (Group 1). Intranasal buserelin spray (150 micrograms x 4 daily) was used in a flare-up protocol (10 women; Group 2) or in a desensitization protocol (10 women; Group 3) in combination with rec FSH. Desensitization using Triptorelin depot (3.75 mgIM) followed by rec FSH was given to 10 women (Group 4), while the same agonist was used subcutaneously (200 micrograms/daily) in Group 5 (10 women). In the five groups rec FSH induced multiple follicular growth. Significantly more ampoules rec FSH were required when a GnRH agonist was used. I all groups 9 to 11 mature oocytes were retrieved per cycle. In 6 couples there was an absence of fertilization due to severe sperm anomalies (5 couples); one couple suffered from idiopathic infertility. Fertilization and cleavage was normal in 44 couples. Forty-three transfers of 2.3 to 2.8 embryos yielded 10 pregnancies. Nine healthy babies were born. Superovulation for IVF was successful and safe by using rec FSH alone or in combination with various GnRHa dosages and protocols.

Buserelin↗
PubMed1991

The correlation between sperm cell morphology and fertilization after zona pellucida slitting in subfertile males.

OBJECTIVE: This work was undertaken to evaluate the correlation between sperm cell morphology and fertilization after zona pellucida slitting in subfertile males. DESIGN: Twenty-two couples who failed at least one in vitro fertilization attempt because of lack of oocytes fertilization underwent a zona-slitting micromanipulative procedure. A total of 245 oocytes were retrieved and inseminated by three different modes: 151 oocytes underwent micromanipulation, 2 were damaged, and the remaining 149 inseminated by the husband's sperm (group A). Fifty-five oocytes were not manipulated and inseminated by the husband's sperm (group B), and 39 oocytes were not manipulated and inseminated by a donor sperm (group C). RESULTS: Fertilization rates were 26.8%, 5.5%, and 53.8% in groups A, B, and C, respectively, and differed significantly between group A and group B. The cleavage rates were lower for oocytes fertilized by the husband's sperm (48.6%) than that obtained by donor (90%), suggesting a sperm factor contributing to this phenomenon. The procedure was most efficient in patients with a total motile sperm count after preparation of greater than or equal to 5 million and with either normal sperm morphology or defects localized to the acrosome or tail region only. Sperm with nuclear morphological abnormalities demonstrated a marked reduction in fertilization potential. CONCLUSION: It is concluded that the zona-slitting technique enhances fertilization of severely subfertile sperm, and its efficacy is affected by sperm morphology and a threshold concentration of motile cells.

Animals↗
PubMed1994

[Comparison of optical analysis and computerized analysis of sperm movement in a program of 316 in vitro fertilizations].

The purpose of this study was to compare visual and computerized assisted semen analysis (CASA) in an in vitro fertilization program. Both methods were performed on the day of 316 oocyte retrievals. CASA was performed using a Hamilton 7.2 device adjusted in accordance with the settings defined by a group of 14 French centers. Despite a few discrepancies, the correlation between visual and CASA sperm counts was significant (p < 0.001), so that between the two total motility assessments. The same correlation (Pearson's test) was observed between the fertilization rate and the parameters obtained by visual analysis and CASA. VAP, VSL, VCL and ALH exhibited threshold values under which the mean fertilization rate was below 50%. This study demonstrates the satisfactory reliability of CASA although it cannot yet replace visual analysis.

Diagnosis, Computer-Assisted↗
PubMed1994

Selective capacity of glass-wool filtration for the separation of human spermatozoa with condensed chromatin: a possible therapeutic modality for male-factor cases?

PURPOSE: The aim of this study was to evaluate chromatin condensation of human spermatozoa following swim-up compared to glass-wool separation. Semen aliquots from men attending an andrological outpatient clinic were processed by means of a swim-up procedure and glass-wool filtration. Chromatin condensation was recorded using aniline blue staining and results were reported according to color intensity of stained sperm heads. Morphometric measurements of sperm heads were performed on stained sperm samples. RESULTS: Glass-wool filtration resulted (i) in a significantly higher total motile sperm count (P < 0.0005) compared to swim-up and (ii) in a significantly higher percentage of normal chromatin-condensed spermatozoa compared to the ejaculate (P < 0.01). CONCLUSION: In contrast, comparing swim-up to the ejaculate, the percentage of matured nuclei (unstained spermatozoa) retrieved following swim-up was significantly lower (P < 0.005). Glass-wool filtration separates human spermatozoa according to motility and size of the sperm head. The size of the sperm head closely correlated with the chromatin condensation quality.

Aniline Compounds↗
PubMed1988

Effect of prostaglandin F(2)alpha at insemination on sperm cell numbers and pregnancy rate in beef cattle.

Fourteen cycling, nonlactating, multiparous beef cows were artificially inseminated (AI) 10 to 12 h after the onset of natural estrus. One unit of frozen-thawed semen containing 100 x 10(6) total sperm cells was deposited into the body of the uterus. Immediately after AI, alternating cows were injected i.m. with either 25 mg (5 ml) of prostaglandin F(2)alpha (PGF) or 5 ml of 0.9% saline-benzyl alcohol control solution. Cows were slaughtered 16 +/- 1 h post AI, oviducts were retrieved, segmented into thirds (upper, middle and lower) and flushed with 1 ml of 0.2% gluteraldehyde in phosphate buffered saline. The number of sperm cells was counted using a phase contrast microscope. There were no right or left side effects (P=0.61) on the number of sperm cells recovered per oviduct within cow (389 vs 553; average SEM = 219). PGF had no effect (P=0.77) on the number of sperm cells recovered per oviduct (642 vs 300; average SEM = 231 for PGF and control females, respectively). More sperm cells were recovered from the lower third segment (P<0.05) compared with the middle and upper segments. Ovulation tended to affect (P=0.10) the number of sperm cells recovered per oviduct (742 vs 200; average SEM = 231). Additionally, 114 beef females (68 Angus x Hereford heifers and 46 Chianina crossbred postpartum suckled cows) were treated as described above following AI at natural estrus with 20 x 10(6) motile sperm cells. Pregnancy rates did not differ significantly in heifers (70.6 vs 58.8%) or in Chianina cows (34.8 vs 52.2%) for control and PGF-treated females, respectively. Overall, pregnancy rates were identical between control and PGF-treated females at 56.1%. In this study, PGF treatment immediately following AI in beef cattle had no effect on the number of sperm cells in the oviducts or on the pregnancy rate.

Journal Article↗
PubMed1998

Does ovarian hyperstimulation syndrome affect the quality of oocytes?

Two wives of a Muslim with severe male factor infertility had simultaneous intracytoplasmic sperm injection (ICSI) treatments. One wife developed ovarian hyperstimulation syndrome (OHSS), and 19 of 27 oocytes retrieved were subjected to ICSI but only one fertilized; the other wife had a normal response to ovarian stimulation, normal fertilization following ICSI, successful treatment and has recently delivered a live-born infant. The wife who suffered from OHSS has since had another ICSI cycle with a normal response to ovarian stimulation, a normal fertilization rate but no pregnancy. The only variable that determined the different rate of fertilization in the simultaneous ICSI cycles appears to be oocyte quality. While the results of frozen embryo replacement cycles following the decision to freeze all embryos following OHSS is generally satisfactory, it is important to counsel couples about the possible detrimental effects of OHSS on oocyte quality.

Adult↗
PubMed1999

Evaluation of the fertilization potential of freshly isolated, in-vitro cultured and cryopreserved human spermatids by injection into hamster oocytes.

The clinical potential for fertilization was examined by using the human sperm-hamster oocyte assay system after microinjection of round (RS), elongating (ES) or elongated (EtedS) spermatids retrieved from obstructive and non-obstructive azoospermic patients. Freshly isolated, in-vitro cultured and cryopreserved spermatids were utilized. For each category of microinjected spermatids, we demonstrated that the more mature the injected spermatid, the higher the incidence of fertilization (for freshly isolated spermatids, P < 0.006 and P < 0.008, for in-vitro cultured spermatids, P < 0.007 and P < 0.007 and for cryopreserved spermatids, P < 0.006 and P < 0.007 for obstructive and non-obstructive azoospermic patients respectively). Short term in-vitro culture of the spermatogenic cells did not improve the incidence of fertilization. However, cryopreservation significantly decreased (P < 0.001) the incidence of fertilization when each corresponding spermatogenic cell stage was compared. The incidence of fertilization was not statistically different when corresponding stages of spermatogenic cells were compared from obstructive and non-obstructive patients.

Animals↗
PubMed1999

Light and electron microscopic analysis of human testicular spermatozoa and spermatids from frozen and thawed testicular biopsies.

The morphological changes caused by freezing and thawing human testicular spermatozoa have been assessed here. Retrieval of testicular biopsies was carried out on six patients with obstructive azoospermia preparatory to intracytoplasmic sperm injection (ICSI). Light microscope analysis was carried out on testicular cells and ultrastructural analysis was carried out on spermatozoa and different spermatid stages before and after the freezing procedure. Upon examination under light microscopy, all germ cells presented increased vacuolization in their cytoplasm and shrinkage or swelling of the nuclei and cytoplasmic membranes. These altered structures were accentuated in the spermatocyte I cell which often presented disrupted membranes. The ultrastructural findings under transmission electron microscopy demonstrated that after freezing and thawing the major types of cryoinjury were the swelling and rupture of inner and outer acrosomal and plasma membranes. The acrosome material often appeared as dispersed material or as condensed spots or was even lost. Such damage was observed mainly at the spermatozoa and late spermatid stages. We conclude that the freezing and thawing of testicular biopsies causes similar morphological damage to testicular spermatozoa and frozen-thawed ejaculated spermatozoa. It is still unclear whether these changes in testicular spermatozoa after freezing and thawing may compromise its use in the ICSI procedure.

Adult↗